JP2007507432A - Process for preparing an oil body comprising an active ingredient - Google Patents

Abstract

The present invention provides a novel emulsion comprising an oil body. The present invention also relates to a novel method for producing a formulation comprising an oil body and an active ingredient, wherein the active ingredient is distributed into the oil body. These methods are particularly useful for producing emulsions with either hydrophobic or amphiphilic biologically active agents.

Description

The present invention provides novel emulsions containing oil bodies. The present invention also relates to a novel method for producing a formulation comprising an oil body and an active ingredient, wherein the active ingredient is distributed into the oil body. These methods are particularly useful for producing emulsions with either hydrophobic or amphiphilic active agents.

Background of the Invention In the seeds of oilseed crops including economically important crops such as soybean, rapeseed, sunflower, safflower, and palm, the water-insoluble oil fraction is an oil body, oleosome, lipid body, or As spherosomes, they are stored in separate subcellular structures that are variously known in the art (Huang 1992, Ann. Rev. Plant Mol. Biol. 43: 177-200). In addition to a mixture of oils (triacylglycerides) that are chemically defined as glycerol esters of fatty acids, oil bodies contain phospholipids and members of related proteins collectively referred to as oil body proteins. From a structural point of view, an oil body is considered to be a triacylglyceride matrix encapsulated by a monolayer of phospholipid in which the oil body is embedded (Huang, 1992, Ann. Rev. Plant Mol. Biol. 43: 177 -200). The seed oil present in the oil body fraction of a plant species is a mixture of various triacylglycerides, the exact composition of which depends on the plant species from which the oil is derived. Manipulating the nature of seed oil and extending it to the naturally available repertoire of plant oil compositions has become possible through a combination of classical breeding and genetic engineering techniques. For an overview of ongoing efforts in this area, see Designer Oil Crops / Breeding, Processing and Biotechnology, edited by DJ Murphy, 1994, VCH Verlagsgesellschaft, Weinheim, Germany.

  Plant seed oil is used in various industrial applications. To obtain the vegetable oils used in these applications, the seeds are crushed or compressed and subsequently purified using processes such as organic extraction, gum removal, neutralization, bleaching, and filtration. Aqueous extraction of vegetable oil seeds has also been demonstrated (eg, Embong and Jelen, 1977, Can. Inst. Food Sci. Technol. J. 10: 239-243). Since the purpose of the process taught by the prior art is to obtain pure oils, oil bodies lose their structural integrity during these manufacturing processes. Thus, prior art emulsions formulated from vegetable oils are generally free of intact oil bodies.

  Voultoury et al. US Pat. No. 5,683,740 and Voultoury et al. US Pat. No. 5,613,583 disclose emulsions containing lipid vesicles prepared from crushed oily plant seeds. In the course of the crushing process described in these patents, the oil bodies substantially lose their structural integrity. Thus, it was disclosed that in the crushing process 70% to 90% of seed oil is released in the form of free oil. Thus, the emulsions that are the subject of these patents are prepared from crushed seeds from which a substantial amount of free oil has been released, but the structural integrity of the oil body is substantially lost. ing. Furthermore, the emulsions disclosed in both of these patents are prepared from a relatively coarse seed extract and contain a number of endogenous seed components, including glycosylated and non-glycosylated oil body proteins. including. The emulsions to which these patents relate include contaminating seed ingredients that impart various undesirable properties to these emulsions that can include allergenic and undesirable odor, flavor, color, and organoleptic properties. Is a disadvantage of these emulsions. Due to the presence of seed contaminants, the emulsions disclosed in these patents have limited use.

  Nondestructive preparation of oil bodies is described in Deckers et al. (US Pat. Nos. 6,183,762, 6,210,742, 6,146,645, 6,372,234, 6,582,710, 6,582,710, 6,596,287, 6,761,914, US 2002/00373036, and US Pat. No. 6,599,513). According to these patents and patent applications, refined preparations of oil bodies are collected as natural emulsions, and further emulsions are used to make the emulsions suitable for cosmetic, pharmaceutical and food applications, among others. In order to achieve the desired emulsification balance, viscosity, stability, and appearance, it can be prepared in the presence of numerous other materials. Additional ingredients to achieve these properties may include water, emulsifiers, stabilizers, thickeners or thinning agents, preservatives, perfumes, or other additives. Of particular interest are oil body preparations containing active ingredients. While simply mixing the assay components of interest with an oil body emulsion can result in an acceptable oil body preparation, it is possible to prepare an oil body emulsion containing an active ingredient that selectively dispenses with the oil body. Particularly desirable. For example, traditionally unstable activity can be stabilized when distributed to the core of the oil body. Furthermore, in oil body formulations used for topical application to human skin, the delivery characteristics of the active ingredient to human skin can be adjusted when the activity is distributed to the oil body. Simply mixing according to the Deckers patent referred to above may facilitate some distribution of the active ingredient, but often, distribution is not achieved or the distribution of the active ingredient is suboptimal. In order to be considered distributed, the activity must be in physical contact with the oil body, for example by bond formation or some other relationship, and must be distributed with the oil body.

  Thus, there is a need in the art to facilitate the distribution of active to intact oil bodies, including, for example, traditionally unstable activity.

SUMMARY OF THE INVENTION The invention referred to herein is of interest because it is distributed in the inner oil core, on the lipid membrane, in the lipid membrane or bound to the outer surface of the lipid membrane of the oil body. Provide one or more hydrophobic and / or amphiphilic activities of the subject.

  A novel system for improved oil body distribution is described herein. This system involves the use of two solvents and the solubilization of activity and activity and a blend of oil body emulsions. This system is more complex than mixing an oil body emulsion with an active ingredient, resulting in an increased activity of partitioning onto or into the oil body. This is particularly useful for solid and semi-solid, hydrophobic and amphiphilic molecules that are particularly difficult to dissolve and often require the use of organic solvents. Removal of the first solvent can be performed in any step if this solvent is not compatible or desirable in the final product. Finally, the activity and solvent are blended and distributed to the oil body.

  The present invention relates to a novel method for producing a formulation comprising an oil body and an active agent, wherein the active agent is distributed into the oil body. Currently, the inventors have discovered a new method for preparing oil bodies containing reactive and unstable active agents in current formulations. Broadly speaking, the present invention provides methods for formulating emulsions containing active agents distributed in oil bodies, where they are stabilized and for topical or oral delivery. Easily available.

Accordingly, the present invention provides a method that provides a method for dispensing an active agent into an oil body, the method comprising:
a) dissolving the active agent in the first solvent;
b) mixing the dissolved active agent with a second solvent; and
c) contacting the solvent mixture with the oil body to distribute the active agent to the oil body.

  In a preferred embodiment of the invention, the active agent is not distributed into the oil body when contacted with the oil body in the absence of a solvent or when the active agent is dissolved in the first solvent. In a further preferred embodiment of the invention, the active agent is selected from the group of active agents consisting of hydrophobic molecules and amphiphilic molecules.

  In a further preferred embodiment of the present invention, the first solvent is an organic solvent. Preferably, the first solvent is substantially removed by evaporation after mixing with the second solvent, or the volume is substantially reduced by dilution.

  In an even more preferred embodiment of the present invention, the second solvent is selected from the group of solvents consisting of water, aqueous buffer, oil, fatty acid, and lipid.

  The method of preparing an oil body comprising the active agent and the resulting emulsion of the present invention can be used in a wide range of applications including in the preparation of personal medical products and dermatological products. Other features and advantages of the present invention will become apparent from the following detailed description. However, the detailed description and specific examples, while indicating preferred embodiments of the invention, are given for purposes of illustration only. This is because various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from this detailed description.

Detailed Description of the Invention As mentioned hereinabove, the present invention relates to a novel method for producing a formulation comprising an oil body and an active agent. The invention also relates to a novel emulsion formulation prepared from oil bodies. Using the present invention, it is possible to distribute the active ingredient, for example, 20% to 30% (dry weight active / dry weight of oil body) to the oil body. The inventors have also steadily produced certain biologically active agents that are reactive and unstable in current formulations, thereby making them more effective as personal medical products and dermatological products. I have found that. The level of achievable activity added, the stability properties of the oil bodies, and the ability to deliver new agents locally make the application to consider of the present invention.

Thus, in accordance with the present invention, a method for dispensing an active agent into an oil body is provided, wherein the method comprises the following steps:
(i) dissolving the active agent in the first solvent;
(ii) mixing the dissolved active agent with a second solvent to obtain a mixture of a first solvent and a second solvent containing the active agent; and
(iii) contacting the active body with the oil body by contacting a mixture of the first solvent and the second solvent with the oil body;

  In preferred embodiments, the active agent does not partition into the oil body when contacted with the oil body in the absence of a solvent or when the active agent is dissolved directly in the first solvent. Preferably, the active agent is further characterized in that the active agent is insoluble in water or essentially insoluble in water. The methods of the invention are particularly useful when the active agent is further insoluble or essentially insoluble in the second solvent.

  The terms “partition”, “partition”, and “distributed” as used herein, activity is localized in the inner oil core, on the lipid membrane, in the lipid membrane. Or bound to the outer surface of the lipid membrane of the oil body.

The solvent term “first solvent” as used herein refers to the first or first solvent used to dissolve the active agent. Preferably, the first solvent is an organic solvent. Examples of organic solvents include, but are not limited to, alcohols, aliphatic hydrocarbons, aromatic hydrocarbons, chlorinated hydrocarbons, glycols, glycol ethers, and their acetates, esters, ethers, and ketones. Examples of alcohols include but are not limited to methanol, ethanol, isopropyl alcohol (isopropanol). Examples of aliphatic hydrocarbons include but are not limited to n-hexane. Examples of aromatic hydrocarbons include but are not limited to toluene, xylene, styrene, and benzene. Examples of chlorinated hydrocarbons include, but are not limited to perchloroethylene, methylene chloride, carbon tetrachloride, methyl chloroform, chloroform, and trichloroethylene. Examples of glycol ethers include butyl cellosolve (2-butoxyethanol), cellosolve (2-ethoxyethanol), methyl cellosolve (2-methoxyethanol), and cellosolve acetate (2-ethoxyethyl acetate). It is not limited to these. Examples of esters include, but are not limited to, methyl formate, ethyl acetate, isopropyl acetate, methyl acetate, secamyl acetate, and isoamyl acetate. Examples of ethers include, but are not limited to, ethyl ether, tetrahydrofuran, dioxane, and isopropyl ether. Examples of ketones include, but are not limited to, acetone, methyl ethyl ketone (MEK), cyclohexanone, and isophorone. More preferably, the first solvent is an alcohol or chlorinated hydrocarbon. Most preferably, the first solvent is selected from the group of solvents consisting of isopropanol, ethanol, and chloroform. Further, the first solvent can be an oil, lipid, or fatty acid.

  The term “second solvent” as used herein refers to a solvent that is mixed with an active agent once it is dissolved in the first solvent. The second solvent can be any solvent that is compatible with the oil body. It should be noted that the method of the present invention is particularly useful when the active agent is not directly soluble in the oil body or the second solvent. The solubility of the active agent in the second solvent includes, but is not limited to, “The CRC, Handbook of Chemistry and Physics” or “The Merck Index”, Merck & Co., Inc. Budavari S (ed.). Can be readily determined by one of ordinary skill in the art by reference to standard chemical indicators that do not provide information regarding stability. The second solvent is selected from the group of solvents consisting of water, aqueous buffer, oil, fatty acid, and lipid. Examples of aqueous buffers include phosphate, phosphate buffered saline, bicarbonate, and buffers containing HEPES (N- [2-hydroxyethyl] piperazine-N '-[2-ethanesulfonic acid]) Liquid, but not limited to. The concentration and pH of the salt may need to be changed to facilitate the distribution of activity, depending on the change in activity. Preferably, the aqueous buffer used is 50 mM sodium phosphate monobasic, pH 8.0 or 25 mM sodium bicarbonate, pH 8.3. Examples of oils include, but are not limited to, oils from the following seeds: rapeseed (Brassica spp.), Soybean (Glycine max), sunflower (Helianthus annuus) , Oil palm (Elaeis guineeis), olive (Olea spp.), Cottonseed (Gossypium spp.), Groundnut (Arachis hypogaea), coconut (Cocus nucifera) )), Castor (Ricinus communis), safflower (Carthamus tinctorius), mustard (Brassica spp.) And Sinapis alba, coriander (Coriandrum sativum) ), Pumpkin (Cucurbita ma xima)), flax / ama (Linum usitatissimum), Brazil nuts (Bertholletia excelsa), jojoba (Simmondsia chinensis), maize (Zea mays), Hana (Crambe abyssinica) and Lucola (Eruca sativa). Other examples of oils include, but are not limited to, synthetic oils, mineral oils, and silicone oils. In a preferred embodiment, the second solvent is safflower oil. The term “fatty acid” is used herein to describe a long hydrocarbon chain terminated by a carboxyl group. Fatty acids are the major components of lipids such as oils, fats, and waxes. Examples of fatty acids include arachidic acid, arachidonic acid, beanic acid, brassic acid, capric acid, caprylic acid, serotic acid, cetrenic acid, erucic acid, gadoleic acid, lauric acid, lauroleic acid, lignoceric acid, linoleic acid, linolenic acid , Margaric acid, merisic acid / triacontanoic acid, myristic acid, montanic acid, myristic acid, oleic acid, palmitic acid, palmitoleic acid, steric acid, ceracoleic acid or nervonic acid, stearic acid It is not limited to. The term “lipid”, as used herein, is a general group of organic substances that are insoluble in polar solvents such as water but are readily soluble in nonpolar organic solvents such as chloroform, ether, benzene. Say. Many, if not all, contain fatty acids as major structural components.

The oil body term “oil body” as used herein means an organelle that stores any subcellular oil or wax. Oil bodies can be obtained from any cell containing oil bodies or oil body-like organelles. This includes animal cells, plant cells, fungal cells, yeast cells (Leber, R. et al., 1994, Yeast 10: 1421-1428), bacterial cells (Pieper-Furst et al., 1994, J. Bacteriol. 176: 4328-4337 ) And algal cells (Roessler, PG, 1988, J. Phycol. (London) 24: 394-400). In a preferred embodiment of the invention, the oil bodies are obtained from plant cells, including pollen, spores, seeds, and cells from the plant's vegetative organ, in which oil bodies or oil body-like organelles are present (Huang, 1992, Ann. Rev. Plant Physiol. 43: 177-200). More preferably, the oil body preparations of the present invention are obtained from plant seeds. Among the plant seeds useful in the present invention, seeds available from plant species selected from the group of plant species consisting of: almonds (Prunus dulcis); anises (Pimpinella anisum) ); Avocado (Persea spp.); Beach nuts (Fagus sylvatica); borage (also known as evening primrose) (Boragio officinalis); Brazil nut (Bertholletia excelsa) Candle nuts (Aleuritis tiglium); calappa (Carapa guineensis); cashew nuts (Ancardium occidentale); castor (Ricinus communis); coconut Cocus nucifera); coriander (Coriandrum sativum); cottonseed (Gossypium spp.); Hamana (Crambe abyssinica); Crepis alpina; croton (croton) (Croton tiglium); Cuphea spp .; Dill (Anethum gravealis); Euphorbia lagascae; Dimorphoteca pluvialis; (Foeniculum vulgaris); groundnuts (Arachis hypogaea); hazelnuts (coryllus avellana); Asa (Cannabis sativa); Ginseng (Lunnaria annua); Jojoba (Simmondsia chinensis); Kapok fruit (Ceiba pentandra); Kukui nuts (Aleuritis moluccana); Amama (Linum usitatissimum); Lupine (Lupinus spp.); Macadamia nut (Macadamia spp.); Corn (Zea mays); Meadowform (Limnanthes alba)); mustard (Brassica spp. and Sinapis alba); olive (Olea spp.); oil palm (Elaeis guineeis); Oitika deer (Licania rigida) Papaya (asimi) Pecans (Juglandaceae spp.); Perilla (Perilla frutescens); Physic nuts (Gatropha curcus); Pyrnuts (Canarium ovatum); Pine Fruit (Pine spp.); Pistachio (Pistachia vera); Pongham (Bongamin glabra); Poppy seed (Papaver soniferum); Rapeseed (Brassica spp.) ); Safflower (Carthamus tinctorius); sesame (Sesamum indicum); soybean (Glycine max); pumpkin (Cucurbita maxima); saraoki (Shorea rubusha) )); Sunflower (Helianthus annuus); Tsukuma (Astocarya spp.); Tungnut (Aleuritis cordata); Mixture of. Most preferably, the plant seed is from a group of plant species including: rapeseed (Brassica spp.), Soybean (Glycine max), sunflower (Helianthus annuus), Oil palm (Elaeis guineeis), cottonseed (Gossypium spp.), Groundnut (Arachis hypogaea), coconut (Cocus nucifera), castor (Ricinus comminis (Ricinus comminis) ), Safflower (Carthamus tinctorius), mustard (Brassica spp.) And Sinapis alba, coriander (Coriandrum sativum), pumpkin (Cucurbita maxima) , Linseed / ama (Rinum Yushitatishimam Linum usitatissimum), Brazil nuts (Bertholletia excelsa), jojoba (Simmondsia chinensis), maize (Zea mays), hamana (Crambe abyssinica) Sativa (Eruca sativa). Oil bodies prepared from safflower (Carthamus tinctorius) are most preferred for use herein,

  In order to prepare oil bodies from plants, such plants can be grown and seeded using agricultural cultivation practices well known to those skilled in the art. Harvesting the seed and, if desired, after removal of substances such as stone or seed hulls (removing the hull), for example by sieving or rinsing the seed and optionally drying. The seed is subsequently processed by mechanical grinding. Preferably, the liquid phase is added prior to seed grinding. This is known as wet grinding. Preferably, the liquid is water, but organic solvents such as ethanol can also be used. Wet milling in the oil extraction process has been reported for seeds from various plant species including: mustard (Aguilar et al. 1991, Journal of Texture studies 22: 59-84), soybean (US Pat. No. 3,971,856; Cater) 1974, J. Am. Oil Chem. Soc. 51: 137-141), peanuts (US Pat. Nos. 4,025,658; 4,362,759), cotton seeds (Lawhon et al., 1977, J. Am. Oil, Chem. Soc. 54: 75-80), and coconut (Kumar et al., 1995, INFORM 6 (11): 1217-1240). It may also be advantageous to immerse the seeds in the liquid phase for a period of about 15 minutes to about 2 days prior to grinding. Soaking can soften the cell walls and facilitate the grinding process. Longer soaking can mimic the germination process and can result in certain advantageous changes in the composition of the seed components.

  The seed is preferably ground using a colloid mill. In addition to colloid mills, other milling and grinding mills capable of processing industrial scale quantities of seeds, including disc mills, colloid mills, pin mills, orbital mills, IKA mills, and industrial scale homogenizers. Devices can also be utilized in the invention described herein. The selection of the mill may depend on the seed throughput requirements as well as the seed source utilized. It is critical that the seed oil bodies remain intact during the grinding process. Therefore, any operating conditions commonly utilized in seed oil processing that tend to destroy the oil bodies are not suitable for use in the subject inventive process. The milling temperature is preferably between 10 ° C and 90 ° C, more preferably between 25 ° C and 50 ° C, and most preferably between 30 ° C and 40 ° C, whereas the pH is Preferably between 2.0 and 11, more preferably between 6.0 and 9.0, and most preferably between 7.0 and 8.5.

  Seed hulls, fibrous material, undissolved carbohydrates, and solid contaminants such as proteins, as well as other insoluble contaminants, are removed from the ground seed fraction. Separation of solid contaminants can be achieved using decantation centrifugation. Depending on the seed throughput requirements, the capacity of the decantation centrifuge can be changed by using other models of decantation centrifuge, such as a three-phase decanter. The operating conditions vary depending on the particular centrifugation that must be utilized and prepared so that the insoluble contaminants settle and remain precipitated during decantation. Partial separation of the oil body phase and the liquid phase can be observed under these conditions.

  After removal of insoluble contaminants, the oil body is separated from the aqueous phase. In one embodiment of the invention, tubular bowl centrifugation is utilized. In a preferred embodiment, disk stack centrifugation is utilized. In other embodiments, hydrocyclones, or phase settling under natural gravity, or any other gravity based separation method may be utilized. It is also possible to separate the oil body fraction from the aqueous phase using size exclusion methods such as filtration, for example membrane ultrafiltration and crossflow filtration methods. An important parameter is the size of the ring dam used to operate the centrifuge. A ring dam is a removable ring with a central annular opening of varying size that regulates the water phase molecules from the oil body and therefore governs the purity of the resulting oil body fraction. The exact ring dam size utilized will depend on the type of centrifuge used, the type of oil seed used, and the desired final consistency of the oil body preparation. In accordance with the present specification, in one embodiment, safflower oil bodies can be obtained using SA-7 (Westphalia) disc stack centrifuge with a ring dam size of 73 mm. The efficiency of the separation is further influenced by the flow rate. In this embodiment, the flow rate is typically maintained between 2.0 l / min and 7.0 l / min and the temperature is preferably maintained between 26 ° C. and 40 ° C. Depending on the model centrifugation used, the flow rate and ring dam size can be adjusted such that optimal separation of the oil body fraction from the aqueous phase is achieved. These adjustments will be readily apparent to those skilled in the art.

  Separation of solids and separation of the aqueous phase from the oil body fraction is also performed simultaneously using a separation method based on gravity, such as a three-phase tubular bowl centrifugation or separation method based on decanter or hydrocyclone or size exclusion. obtain.

  The composition obtained at this stage in the process is generally relatively coarse and includes glycosylated and unglycosylated proteins and other contaminants such as glucocinylate or its degradation products. Contains a large number of seed proteins. In a preferred embodiment of the invention, a significant amount of seed contaminants is removed. In order to achieve removal of contaminating seed material, the oil body preparation obtained upon separation from the aqueous phase can be resuspended and the resuspended fraction can be centrifuged. Washed at least once. This process results in what is called a washed oil body preparation for purposes of this application. The number of washes generally depends on the desired purity of the oil body fraction. Depending on the washing conditions utilized, an essentially pure oil body preparation can be obtained. In such a preparation, the only protein present is an oil body protein. Tubular bowl centrifugation or other centrifugation, such as hydrocyclone or disc stack centrifugation, can be used to wash the oil body fraction. The oil body wash can be performed in water, a buffer system, eg, sodium chloride at a concentration between 0.01M and at least 2M, 0.1M sodium carbonate at high pH (11-12), a low salt buffer, eg, 50 mM Tris- It can be carried out using HCl pH 7.5, organic solvents, surfactants, or any other liquid phase. In embodiments where high purity oil bodies are deemed desirable, washing is preferably performed at high pH (11-12). The liquid phase used for washing, and the washing conditions, such as pH and temperature, can be varied depending on the type of seed used. Washing at a number of different pHs between pH 2 and pH 11-12 can be beneficial when this allows stepwise removal of contaminants in a particular protein. The washing conditions are selected so that the washing process results in a significant amount of contaminant removal without compromising the structural integrity of the oil body. In embodiments where more than one cleaning step is performed, the cleaning conditions can vary for different cleaning steps. SDS gel electrophoresis or other analytical techniques can be conveniently used to monitor the removal of seed proteins and other contaminants during oil body washing. It is not necessary to remove all of the aqueous phase between washing steps, and the final washed oil body preparation will contain water, a buffer system, e.g. 50 mM Tris-HCl pH 7.5, or any other liquid phase. If so desired, the pH may be adjusted to any pH between pH 2.0 and 11, more preferably between 6.0 and 9.0, and most preferably between 7.0 and 8.5. obtain.

  The process for producing the oil body preparation may be performed in a batch operation or a continuous flow process. Especially when a disk stack is used, the pump system is conveniently set up to produce a continuous flow. This pump can be, for example, an air operated double diaphragm pump, a hydraulic pump, a positive displacement pump, or a peristaltic pump. In order to maintain a homogeneous consistency supply for decantation centrifugation and annular bowl centrifugation, for example, an IKA homogenizer or the like can be added during the separation process. An in-line homogenizer can also be added between the various centrifuge or size exclusion based devices utilized to wash the oil body preparation. Ring dam size, buffer composition, temperature, and pH may be different in each wash step from the ring dam size utilized in the first separation step.

Activity In accordance with the present invention, a wide variety of biologically active ingredients can be formulated with the oil bodies of the present invention. The terms “biologically active agent”, “active”, “active agent”, and “active ingredient” as used herein refer to any physiology when administered to a living organism. Means any agent that has a detectable biological effect, including clinical, diagnostic, prophylactic, or pharmacological effects. These terms are meant to include, but are not limited to, any pharmacological, therapeutic, nutritional, dermatological, or medicated cosmetic agent. Furthermore, the terms “biologically active agent”, “active”, “active agent”, and “active ingredient” as used herein are preferably oil bodies, water, aqueous solutions, oils, Fatty acid, or a compound that is directly insoluble in lipids, but whose activity is soluble in organic solvents. The activity may be capable of enhancing or improving the physical appearance, health, adaptability, or ability of surface areas of the human body, eg, skin, hair, scalp, teeth, and nails. The activity can be up to a clinically significant level, up to some level that can be added in excess (20% -30% dry weight activity / oil body dry weight). The amount of activity formulated will depend on the desired effect and the activity selected. In general, the amount of activity ranges from 0.0001% (w / w) to about 50% (w / w). More preferably, however, the amount of activity in the final composition is from 0.01% (w / w) to about 20% (w / w), most preferably from 0.1% (w / w) to about 10% ( w / w). Depending on the chemistry of the activity, the activity can be incorporated into the final formulation in a variety of ways, for example, the amphiphilic activity can be partitioned into the phospholipid membrane of the oil body, while the hydrophobic Sexual activity can be distributed to the lipid core of the oil body.

  The active agent is dissolved in a first solvent, which is an amount of the first solvent that is sufficient to dissolve the active agent. The amount will vary depending on the activity and solvent. The amount can be readily determined by one skilled in the art. Standard chemistry that provides information on stability, including but not limited to “The CRC, Handbook of Chemistry and Physics” or “The Merck Index”, Merck & Co., Inc. Budavari S By referring to the relevant indicators. Once the activity is dissolved in the first solvent, the dissolved activity is mixed with the second solvent.

  Preferably, the first solvent is substantially removed after mixing with the second solvent. In certain embodiments, the first solvent is substantially removed by evaporation or the volume is substantially reduced by dilution. Examples of methods for evaporating the first solvent include, but are not limited to, exposing the sample to a stream of either compressed air, oxygen, or nitrogen. Preferably, the sample is exposed to a stream of nitrogen. The term “substantially removed” as used herein preferably removes about 90% to 99.9% of the first solvent, more preferably about 95% to about 99.9%. Of the first solvent is removed, and most preferably, from about 99% to about 99.9% of the first solvent is removed.

  The oil body is incubated with a solvent / activity mixture to facilitate the distribution of activity to the oil body. This incubation is preferably carried out at temperatures above 0 ° C. and can be carried out at room temperature or at elevated temperatures. This incubation can be performed overnight, for example, at a temperature between 34 ° C. and 37 ° C. to ensure optimal distribution.

  Hexane extraction can be used to measure the amount of activity present in free oil (free oil is defined as oil not contained in the oil body. Intact oil body is due to hexane alone. Note that it is largely resistant to extraction) (Tzen et al. (1997) J. Biochem 121: 762-768.). Once the free oil is removed from the active / oil body fraction, analysis of the total remaining oil can be performed using, for example, hexane: isopropanol, chloroform, or chloroform: methanol solution extraction. The amount of activity present in both the free oil fraction and the total oil fraction includes, but is not limited to, high performance liquid chromatography (HPLC), spectrophotometry, fluorescence, or activity assay, depending on the activity. It can be measured by a number of methods. By comparing the amount of activity in both the free oil fraction and the total oil fraction, the average amount of activity incorporated into the intact oil body can be determined. In general, the efficiency of distribution of activity to intact oil bodies can range from about 10% to about 99.9%. More typically, however, the efficiency of partitioning activity into intact oil bodies ranges from about 50% to about 99%, and most typically from about 90% to about 99%.

The term “hydrophobic” as used herein refers to a substance that is not readily soluble in polar solvents such as water. In general, as the hydrophobicity increases, the tendency of the material to partition into the nonpolar solvent increases. The hydrophobic nature of a molecule can be measured by the partition coefficient of the molecule. Simply put, the partition coefficient is the ratio of the equilibrium concentration between the two immiscible phases in contact. For example, the octanol / water partition coefficient (K _OW , P _OW , or P value) is _{calculated as} that in the octanol (nonpolar) phase versus the concentration of the chemical / activity in the water (polar) phase of the two-phase octanol / water system. Concentration ratio. Compounds with high P values are considered relatively hydrophobic. Since measured values range from <10 ⁻⁴ to> 10 ⁺⁸ (at least 12 orders of magnitude), logarithm (log P) is commonly used to characterize that value. The logP values are, for example, using reverse phase HPLC (Yamagami and Haraguchi, 2000. Chem Pharm Bull (Tokyo) 48 (12): 1973-7) or by Syracuse Research Corporation using atom / fragment contribution methods. It can be determined empirically by using a computer program such as the developed KowWin program.

  In preferred embodiments, the log P value of the active compound ranges from about 0 to about 8. In a more preferred embodiment, the log P value ranges from about 2 to about 7. In the most preferred embodiment, the log P value ranges from about 3 to about 7.

  One particularly preferred hydrophobic activity that can be used according to the present invention is clobetasol propionate. Other synonyms and derivatives for clobetasol propionate include clobetasol, clobetasol 17-propionate, (11β, 16β) -21-chloro-9-fluoro-11-hydroxy-16-methyl-17- (1-oxopropoxy ) Pregna-1,4-diene-3,20-dione, Dermoval, Dermovate, Dermoxin, Dermoxinale, Temovate, and derivatives thereof It is not limited to these. Conditions that clobetasol propionate can be used to treat include inflammatory and pruritic symptoms of moderate to severe corticosteroid responsive skin disease. Examples of these signs include allergic reaction, atopic dermatitis, contact dermatitis, eczema, lichen planus, sclerotrophic lichen, phimosis, pruritus, psoriasis, scalp skin disease, seborrheic skin Including, but not limited to, inflammation and skin irritation.

  Another particularly preferred hydrophobic activity that can be used according to the present invention is diclofenac. Other synonyms and derivatives for diclofenac include 2-[(2,6-dichlorophenyl-amino] benzeneacetic acid, [o-2,6-dichloroanilino) phenyl] -acetic acid, voltarol, catafram ( Catafram (Diclofenac potassium), Blotaren (Diclofenac sodium), Brotalene-XR, Solaraze, Allvoran, Benfofen, Dealgic, Deflamat, Delphinac, Delphinac Diclomax, Miclometin, Diclophlogont, Diclo-Puren, Dicloreum, Diclo-Spondyril, Delobasan, Duravolten, Ecofenac, Effekton, Lexobene, Motifene, Neriodin, Novapirina, Primofenac, Prophenatin, Rewodina, Rhumalgan, Trabona, Tudomin, Tudomin These include, but are not limited to, Valetan, Voldal, Xenid, and derivatives thereof. Diclofenac is an anti-inflammatory anagenic that is effective in treating heat, pain, and inflammation throughout the body. Conditions that diclofenac can be used to treat include pain caused by arthritis and gout, tenderness, reduction of inflammation (swelling) and stiffness, menstrual pain and relief after surgery or postpartum, rheumatoid arthritis, osteoarthritis, This includes, but is not limited to, ankylosing spondylitis, cataract or corneal reaction surgery and post-operative inflammation after UV keratitis.

  Yet another particularly preferred hydrophobic activity that can be used according to the present invention is dithranol. Other synonyms and derivatives for dithranol include 1,8-dihydroxy-9 (lOH) -anthracenone, 1,8-dihydroxyanthrone, anthralin, Anthraforte, Anthranol, Anthrascalp ), Antraderm, Cignolin, Dithrocream (registered trademark), Drithrocreme, Dithro-Scalp, Micanol, Psoradrate, Procidarm (Prosoderm) Prosiderm), sorin (Psorin), and derivatives thereof. Symptoms where disranol can be used to treat include, but are not limited to, subacute and chronic psoriasis.

  One particularly preferred hydrophobic activity that can be used according to the present invention is retinoic acid. Other synonyms and derivatives for retinoic acid include (all-E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexen-1-yl) -2,4,6 , 8-Nonatetraenoic acid, vitamin A acid, tretinoin, Aberel, Airol, Avita, Epi-Aberel, Eudyna, Kerlocal, Renova ) (Trademark), Retin-A (trademark), retinol, Vesanoic and derivatives thereof. Symptoms where retinoic acid can be used to treat include mild to moderate acne and sun-damaged (light-aged) skin (ie wrinkles, mottled hyperpigmentation, and excess to sunlight) Reducing skin roughness associated with exposure), but is not limited thereto.

  Another particularly preferred hydrophobic activity that can be used according to the present invention is lidocaine. Other synonyms and derivatives for lidocaine include 2-diethylamino-2 ', 6'-acetoxylidide, 2- (diethylamino) -N- (2,6-dimethylphenyl) acetamide, Anestacon, quivasar (Cuivasal), Duncaine, EMLA (R), Gravocain, Isicaine, Leostesin, Lidocaine, Lidocaine Base and Lidocaine HCl USP (LIDOCAINE BASE AND LIDOCAINE HCl USP) (Lidothesin), Lignocaine, Rucaina, Silestesin, ω-diethylamino-2 ', 6'-dimethylacetanilide, Xylestesin, Xylocaine, Xylocitin, Xylocitin, Xylotox) and derivatives thereof, but are not limited to these. Symptoms for which lidocaine can be used to treat include, but are not limited to, ventricular arrhythmias, ventricular extrasystoles (PVC), tachycardia, and fibrillation.

  Yet another particularly preferred hydrophobic activity that can be used according to the present invention is clindamycin. Other synonyms and derivatives for clindamycin include 7-deoxy-7 (S) -chlorolincomycin, 7 (S) -chloro-7-deoxylincomycin, Antirobe, cleosin , Clindamycin, Clindatech, Dalacin, Daracin C, Dalactine, Hydrochloride monohydrate (Daractin), Klimicin, Klimycin C, Krimycin methylxamide)- 1-thio-L-threo-α-D-galacto-octylpyranoside, L-threo-α-D-galacto-octopyranoside, methyl 7-chloro-6,7,8-trideoxy-6-(((l -Methyl-4-propyl-2-pyrrolidinyl) carbon yl) amino) -1-thio-, (2S-trans)-, hydrochloride monohydrate, methyl 7-chloro-6,7,8-trideoxy-6- (l-Methyl-trans-4-propyl-L-2-pyro Gincarboxamide) -1-thio-L-threo-α-D-galacto-octopyranoside, methyl 7-chloro-6,7, thio (hio) -L-threo-α-D-galacto-octopyranoside, Sobelin , U-21251, and derivatives thereof, but are not limited to these. Symptoms for which clindamycin may be used to treat include, but are not limited to, bacteroides fragilis infection in infants.

  Yet another particularly preferred hydrophobic activity that can be used according to the present invention is benzoyl peroxide. Other synonyms and derivatives for benzoyl peroxide include 2,3,6-TBA, acetoxyl, incidol, loroxide, lucidol, luperco, luperox ) Fl, nayper b and bo, Nericur, Norox bzp-250, Norox bzp-c-35, Novadelox, Oxy-10, OXY-5, Oxy-5, Oxy 10 , Oxylite, oxy wash, PanOxyl, Peroxydex, Persadox, Persa-gel, quinolor compounds, sanoxit, Superox, TCBA, Theraderm, Topex, Tribac, vanoxide, Xerac, Zerac BP 1 0, Zerak BP 5, Acnegel, Aztec bpo, Benoxyl, Benzac, Benzagel 10, Benzaknen, Benzaknew, benzoic acid, peroxide , Benzoperoxide, benzoyl peroxide, benzoyl peroxide, residual water, benzoyl peroxide, dibenzoyl peroxide, diphenylglyoxal peroxide, BPO, BZF-60, Cadet, cadox bs, Desanden ), Debroxide, dry and clear, epi-clear, fostex, garox, and derivatives thereof. Symptoms where benzoyl peroxide can be used to treat include, but are not limited to, mild to moderate acne.

  Yet another particularly preferred hydrophobic activity that can be used according to the present invention is cyclosporin A. Other synonyms and derivatives for cyclosporin A include the antibiotic s7481fl, cyclosporin (cyclosporin A, cyclosporin A, ol 27-400, ramifhin A, s7481fl, and derivatives to these Symptoms that can be used to treat cyclosporin A include, but are not limited to, patients receiving organ (eg, kidney, liver, and heart) transplants, psoriasis, and the body's natural immunity in rheumatoid arthritis Is included, but is not limited thereto.

  In another embodiment, the anticancer drug doxorubicin (also known as adriamycin) can be used and formulated as an oil body emulsion, according to the present description.

The term “amphiphilic” or “amphipathic” as used herein refers to molecules having two distinguishable elements that differ in their affinity for solutes and solvents. One part of the molecule has affinity for polar solvents such as water and is said to be hydrophilic. The second part of the molecule has an affinity for non-polar solvents such as hydrocarbons, which are said to be hydrophobic. Amphiphilic molecules exhibit unique behavior when interacting with water, where the polar or hydrophilic portion of the molecule “pursues” to interact with water, whereas nonpolar Or the hydrophobic part “blocks” the interaction with water. The balance between the hydrophilic and hydrophobic parts in the amphiphilic molecule is used as a classification method (hydrophilic-lipophilic balance, HLB). HLB values for commonly used amphiphilic molecules are readily available in the literature (eg, “Handbook of Pharmaceutical Excipients”, The Pharmaceutical Press. London, 1994). The HLB system was originally devised by Griffin (J. Soc. Cosmetic Chem., 1, 311, 1949). Griffin defines the HLB value of an amphiphilic molecule as the mole percent of hydrophilic groups divided by 5, where a completely hydrophilic molecule (without a non-polar group) has an HLB value of 20. Had. This simple approach to calculate HLB values is only applicable to polyoxyethylene ethers. Consequently, for other amphiphilic molecules, HLB values have been derived from a variety of properties such as water solubility, dielectric constant, interfacial tension, and cloud point. Such dispersants preferably have a hydrophilic-lipophilic balance between 1 and 20 (Griffin, WC, J. Soc. Cos. Chem. 1, 1949. 311: J. Soc. Cos. Chem 5, HLB values as defined in 1954, 249). Davis et al. Proc. 2ns Int. Cong. Surface Act. Vol 1 Butterworths, 1959, London proposed a more general empirical formula that associates constants for different hydrophilic and hydrophobic groups:
HLB = [(n _H × H) -n _L × L] +7
Here, H and L are constants assigned to the hydrophilic group and the hydrophobic group, respectively, and n _H and n _L are the number of these groups per molecule. For the purposes of the present invention, HLB is an empirical amount on an arbitrary scale, which is a measure of the polarity of an amphiphilic molecule or mixture of amphiphilic molecules. See P. Becher et al., “Nonionic Surfactant, Physical Chemistry” Marcel Dekker, NY (1987), pages 439-456. Preferably, in accordance with the present invention, the amphiphilic activity HLB value ranges from about 1 to about 14, more preferably the active HLB ranges from about 4 to about 10, and most preferably the activity. The HLB value is in the range of about 6-8.

  One particularly preferred amphipathic activity that can be used according to the present invention is amphotericin B. Other synonyms and derivatives for amphotericin B include, but are not limited to, amphotericin B deoxycholic acid, fandizone ™, and derivatives thereto. Symptoms where amphotericin B can be used to treat include, but are not limited to, fungal infections.

  One particularly preferred amphiphilic activity that can be used according to the present invention is phosphatidylcholine. Other synonyms and derivatives for phosphatidylcholine include, but are not limited to lecithin and its derivatives. Phosphatidylcholine is a membrane phospholipid. Phosphatidylcholine has been found in many skin care products and has been used in cosmetic surgery (ie, injected into a fat pad under the eye to minimize swelling).

  One particularly preferred amphiphilic activity that can be used according to the present invention is tetracaine. Other synonyms and derivatives for tetracaine include amethocaine, 2-dimethylaminoethyl, 4- (butylamino) benzoic acid 2- (dimethylamino) ethyl ether monohydrochloride, 4- (butylamino) benzoic acid, Anethaine, Butethanol, Tonexol, 4- (Butylamino) -benzoic acid, 2- (Dimethylamino) ethyl ester, Dicain, Decicain, Pontocaine, and These derivatives are included, but are not limited to these. Tetracine is an effective local anesthetic for topical application. Examples of these topical applications include, but are not limited to, venepuncture or venous cannulation and anesthesia prior to minor eye surgery.

  One particularly preferred amphiphilic activity that can be used according to the present invention is actinomycin. Other synonyms and derivatives for actinomycin include 3H-phenoxazine-1,9-dicarboxamide, 2-amino-N, N′-bis [hexadecahydro-2,5,9-trimethyl-6, 13-bis (1-methylethyl) -1,4,7,11,14-pentaoxo-lH-pyrrolo [2,1-i] [1,4,7,10,13] oxatetraazacyclohexadecin- 10-yl] -4,6-dimethyl-3-oxo-, ACT, hbf 386, Lyovac cosmegen, meractinomycin, oncostatin K, X 97, actinomycin A IV, Actinomycin 7, actinomycin AIV, actinomycin dioic D acid, dilactone, actinomycin C1, actinomycin cl, actinomycin D, (-)-actinomycin d, actinomycin i, actinomycin Il, actinomycin IV, actinomycin Mycin- [threo-val-pro-sar-meval], actinoma Syn X 1, actinomycin xi, actinomyein-theo-val-pro-sar-meval, ACTO-D, AD, dilactone actinomycin dioic D acid, dilactone actinomycin D acid, C1, cosmegen (Cosmegen), dactinomycin, dactinomycin D, dactinomyein d, and derivatives thereof, including but not limited to. Symptoms where actinomycin D can be used to treat include, but are not limited to, cancer and are used as antibiotics.

Further activities intended for use in the compositions described herein include the following categories and examples of activities and alternative types of these activities, such as alternative salt forms, Free acid forms, free base forms, and hydrates include:
Analgesics / antipyretics (e.g. aspirin, acetaminophen, ibuprofen, naproxen sodium, buprenorphine, propoxyphene hydrochloride, propoxyphene napsilate, meperidine hydrochloride, hydromorphone hydrochloride, morphine, oxycodone, codeine, dihydrocodeine hydrogen tartrate, pentazocine, Hydrocodone bitartrate, levorphanol, diflunisal, trolamine salicylate, nalbufen hydrochloride, mefenamic acid, butorphanol, choline salicylate, butarbital, phenyltroxamine citrate, diphenhydramine citrate, methotrimeprazine, cinnamedrine hydrochloride Salt, and meprobamate);
• Antiasthamatics (eg, ketotifen and traxanox);
Antibiotics (eg neomycin, streptomycin, chloramphenicol, cephalosporin, ampicillin, penicillin, tetracycline, and ciprofloxacin);
Antidepressants (e.g. nefopam, oxypertin, doxepin, amoxapine, trazodone, amitriptyline, maprotiline, phenelzine, desipramine, nortriptyline, tranylcypromine, fluoxetine, doxepin, imipramine, imipramine pamoate, isocarboxazide, and promibramine Triptyline);
Anti-diabetic agents (eg biguanides and sulfonylurea derivatives);
Antifungal agents (eg griseofulvin, ketoconazole, itraconazole, amphotericin B, nystatin, and candicidin);
Antihypertensive agents (e.g., propanolol, propaphenone, oxyprenolol, nifedipine, reserpine, trimethaphan, phenoxybenzamine, pargyline hydrochloride, deserpidine, diazoxide, guanethidine monosulfate, minoxidil, resinnamine, nitroprusside sodium, Low orfia serpentina, arceloxylone, and phentolamine);
Anti-inflammatory agents (e.g. (non-steroidal) indomethacin, ketoprofen, flurbiprofen, naproxen, ibuprofen, ramifenazone, piroxicam, biphenylcarboxylic acid derivatives, acetominaphen, (steroidal) hydrocortisone, cortisone, dexamethasone, fluazacoat, Celecoxib, rofecoxib, hydrocortisone, prednisolone, and prednisone);
Antineoplastic agents (e.g., cyclophosphamide, actinomycin, bleomycin, daunorubicin, doxorubicin, epirubicin, mitomycin, methotrexate, fluorouracil, carboplatin, carmustine (BCNU), methyl-CCNU, cisplatin, etoposide, camptothecin and its derivatives, Phenesterine, paclitaxel and its derivatives, docetaxel and its derivatives, vinblastine, vincristine, tamoxifen, and pipersulfan);
Anxiolytics (eg, lorazepam, buspirone, prazepam, chlordiazepoxide, oxazepam, dipotassium chlorazepate, diazepam, hydroxyzine pamoate, hydroxyzine hydrochloride, alprazolam, droperidol, halazepam, chlormezanone, and dantrolene);
Immunosuppressants (eg cyclosporine, azathioprine, mizoribine, and FK506 (tacrolimus));
Anti-migraine drugs (eg, ergotamine, propanolol, isometheptene mucate, and dichlorarfenazone);
Sedative / hypnotics (eg, barbituric acid, eg, pentobarbital, and secobarbital; and benzodiazapine, eg, flurazepam hydrochloride, trizolam, and midazolam);
Anti-angina agents (eg β-adrenergic blockers; calcium channel blockers such as nifedipine and diltiazem; and nitrate compounds such as nitroglycerin, isosorbide dinitrate, pentaerythritol tetranitrate, and erythrityl tetranitrate);
Antipsychotics (e.g. haloperidol, loxapine succinate, loxapine hydrochloride, thioridazine, thioridazine hydrochloride, thiothixene, fluphenazine, fluphenazine decanoate, fluphenazine enanthate, trifluoperazine, chlorpromazine, perphenazine, lithium citrate, And prochlorperazine);
An antidepressant (eg lithium carbonate);
Antiarrhythmic drugs (for example, bretylium tosylate, esmolol, verapamil, amiodarone, encainide, digoxin, digitoxin, mexiletine, disopyramide phosphate, procainamide, quinidine sulfate, quinidine gluconate, quinidine polygalacturonic acid, flecainide acetate, Tocainide and lidocaine);
Anti-arthritic agents (eg phenylbutazone, sulindac, penicillamine, salsalate, piroxicam, azathioprine, indomethacin, meclofenamic acid, gold sodium thiomalate, ketoprofen, auranofin, aurothioglucose and tolmethine sodium);
Anti-gout drugs (eg colchicine and allopurinol);
• anticoagulants (eg, heparin, heparin sodium, and warfarin sodium);
Thrombolytic agents (eg urokinase, streptokinase and alteplase);
An antifibrinolytic agent (eg aminocaproic acid);
Blood rheology agents (eg pentoxifylline);
Antiplatelet drugs (eg aspirin);
Anticonvulsants (eg, valproic acid, divalproex sodium, phenytoin, phenytoin sodium, clonazepam, primidone, phenobarbitol, carbamazepine, amobarbital sodium, methosuximide, metalbital, mehobarbital, mephenytoin, fenceximide , Parameterdione, ethotoin, phenacemide, sodium secobarbitol, dipotassium chlorazepate, and trimetadione);
Antiparkinsonian drugs (eg ethosuximide);
Antihistamines / antipruritics (e.g. hydroxyzine, diphenhydramine, chlorpheniramine, bromophenylamine maleate, cyproheptadine hydrochloride, terfenadine, clemastine fumarate, triprolidine, carbinoxamine, diphenylpyralin, phenindamine, azetazine, tripelenamine, dexchlorphenylamine Maleate, methodirazine, and);
• Agents useful for calcium regulation (eg, calcitonin and parathyroid hormone);
Antibacterial agents (for example, amikacin sulfate, aztreonam, chloramphenicol, chloramphenicol palmitate, ciprofloxacin, clindamycin, clindamycin palmitate, clindamycin phosphate, metronidazole, metronidazole hydrochloride, Gentamicin sulfate, lincomycin hydrochloride, tobramycin sulfate, vancomycin hydrochloride, polymyxin B sulfate, sodium colistimate, and colistin sulfate);
Antiviral agents (eg interferon α, β or γ, zidovudine, amantadine hydrochloride, ribavirin, and acyclovir);
An antimicrobial agent (e.g. cefasporin, e.g. cefazolin sodium, cefadine, cefaclor, cefapirin sodium, ceftizoxime sodium, cefoperazone sodium, cefotetan disodium, cefuroxime e azotyl, cefotaxime sodium, cefadroxyl monohydrate, cephalexin, Cephalothin sodium, cephalexin hydrochloride monohydrate, cefamandole nafate, cefoxitin sodium, cefoniside sodium, ceforanide, ceftriazone sodium, ceftazidine, cefadroxyl, cephalazine, and cefuroxime sodium; penicillin such as ampicillin , Amoxicillin, penicillin G benzathine, cyclacillin, ampicillin sodium, penicillin G potassium Penicillin V potassium, piperacillin sodium, oxacillin sodium, bacuanpicillin hydrochloride, cloxacillin sodium, ticarcillin disodium, azulocillin sodium, carbenicillin indanyl sodium, penicillin G procaine, methicillin sodium, and naphthillin sodium; erythromycin, for example Erythromycin ethyl succinate, erythromycin, erythromycin estrate, erythromycin lactobionate, erythromycin stearate, and erythromycin ethyl succinate; and tetracyclines such as tetracycline hydrochloride, doxycycline hykrate, minocycline hydrochloride, azithromycin, triclothromycin , Tornaf Benzoate, chlorhexidine, benzoyl peroxide)
An anti-infective drug (eg GM-CSF);
Bronchodilators (eg, sympathomimetic agents, eg epinephrine hydrochloride, metaproterenol sulfate, terbutaline sulfate, isoetarine, isoetarine mesylate, isoetarine hydrochloride, albuterol sulfate, albuterol, vitorterol mesylate, isoproterenol hydrochloride Terbutaline sulfate, epinephrine hydrogen tartrate, metaproterenol sulfate, epinephrine sulfate, and epinephrine hydrogen tartrate; anticholinergics such as ipratropium bromine; xanthines such as aminophylline, diphylline, metaproterenol sulfate, and aminophylline such as cromolyn sodium Mast cell stabilizers; inhalant corticosteroids such as beclomethasone dipropionate (BDP) and beclomethasone dipropionate monohydrate; salbutamol; Of ipratropium; budesonide; ketotifen; salmeterol; Kishinafote (xinafoate); terbutaline sulfate; triamcinolone; theophylline; nedocromil sodium; metaproterenol sulfate; albuterol; flunisolide; fluticasone propionate)
Steroidal compounds and hormones (eg androgens such as danazol, testosterone cypionate, fluoxymesterone, ethyl testosterone, testosterone enate, methyltestosterone, fluoxymesterone and testosterone cypionate; estrogens such as estradiol Progestins such as methoxyprogesterone acetate and norethindrone acetate; corticosteroids such as triamcinolone, betamethasone, sodium betamethasone phosphate, dexamethasone, dexamethasone sodium phosphate, dexamethasone prednisone acetate, methylprednisolone acetate Turbid, triamcinolone acetonide, methylprednisolone, lithium Sodium prednisolone, sodium methylprednisolone succinate, hydrocortisone sodium succinate, triamcinolone hexaacetonide, hydrocortisone, hydrocortisone cypionate, prednisolone acetate, fludrocortisone acetate, parameterzone acetate, prednisolone tebutate, prednisolone acetate, sodium prednisolone phosphate And sodium succinate hydrocortisone; thyroid hormones such as levothyroxine sodium);
Hypoglycemic agents (eg human insulin, purified bovine insulin, purified porcine insulin, glyburide, chlorpropamide, glipizide, tolbutamide, and tolazamide);
Lipid-lowering agents (eg clofibrate, dextrothyroxine sodium, probucol, pravastatin, atorvastatin, lovastatin, and niacin);
A protein (eg, DNase, arginase, superoxide dismutase, and lipase); a nucleic acid (eg, sense or antisense encoding any therapeutically useful protein, including any protein described herein) Nucleic acid);
• agents that are useful for stimulating erythropoiesis (eg erythropoietin);
An anti-ulcer / anti-reflux agent (eg, famotidine, cimetidine, and ranitidine hydrochloride);
Antiemetics / emetics (eg, meclizine hydrochloride, nabilone, prochlorperazine, dimenhydrinate, promethazine hydrochloride, triethylperazine, and scopolamine);
Fat-soluble vitamins (eg, vitamins A, D, E, K, etc.); and other drugs such as mitotane, halonitrosourea, anthrocycline, and ellipsthecin;
Sunscreen activity (eg, para-aminobenzoic acid (PABA), octyl salicylate, octyl methoxycinnamate (Parasol MCX); titanium dioxide)
・ Wrinkle remover and anti-aging activity (eg retinoic acid)
Whitening and bleaching activity (eg ascorbyl palmitate and licorice root)
Anti-acne activity (eg hydrocortisone and benzoyl peroxide)
Vitamin activity (eg vitamin A and derivatives including retinoic acid, retinyl aldehyde, retin A, retinyl palmitate, adapalene, and β-carotene; vitamin D including calcipotriene (vitamin D3 analog); Vitamin E including the components α-, β-, γ-, δ-tocopherol and cotrienol and mixtures thereof and vitamin E derivatives including vitamin E palmitate, vitamin E linoleate, and vitamin E acetate; vitamin K and derivatives)
Lipids (for example, triacylglycerol; fatty acids such as γ-linolenic acid; waxes; cholesterol; sphingolipids; ceramides; phospholipids; and mixtures thereof)
・ Dyes (for example, titanium dioxide, zinc oxide, zirconium dioxide, methozalene, trioxalene, carotenoids (α-carotene, β-carotene, lutein, lycopene), chlorophyll (a and b), xanthophylls)
Antioxidants (eg vitamin E and vitamin E mimics, alpha lipoic acid, coenzyme Q (CoQ, Q, ubiquinone), vitamin A, carotene, lycopene, lipoic acid, melatonin, some polyphenols, some flavonoids )
-Antifungal agents (eg, rilopirox, lanoconazole, benzylamine derivatives, imadozole, amphotericin B)
Perfumes (for example, acetanisole, acetophenone, acetyl cedrene, methylnonyl acetaldehyde, heliotropin, sandera, methoxycitranelar, hydroxycitranelar, geraniol, benzaldehyde, linalool, p-tert-butylcyclohexyl acetate, cinnamyl acetate 1-menthol, vanillin, sandlewood oil, angelic root oil, bergamot oil, butterleaf oil, cassia oil, chamomile oil, lemon oil, lavender oil, ylang ylang oil)
Insect repellents (eg N, N-diethyl-3methylbenzamide, N, N-diethyl-m toluamide (DEET), dimethyl phthalate (DMP), citronella or eucalyptus oil, pyrethrin, pyrethroid ).

  A description of these and other classes of useful activity, as well as a list of species within each class, can be found in “Martindale, The Extra Pharmacopoeia”, 30th Edition (The Pharmaceutical Press, London 1993). This disclosure is incorporated herein by reference in its entirety.

  The following non-limiting examples are illustrative herein.

Example <br/> Example 1
Obtaining a washed oil body preparation from safflower This example illustrates the recovery of an oil body fraction from safflower. The resulting preparation contains an intact washed oil body.

Seed decontamination A total of 45 kg of dried safflower (Carthamus tinctorius) seeds were used twice using approximately 120 L of tap water at 65 ° C and 1 using approximately 120 L of tap water at approximately 15 ° C. Washed twice. Washing was carried out in one barrel with a sieve to separate the water to be discarded.

Seed grinding MZ-120 rotor / stator grinding set with crossed teeth and washed seeds while supplying approximately 100 L of pH 7.0 25 mM NaH ₂ P0 ₄ buffer through an externally connected hose prior to grinding It was poured through a hopper of a colloid mill (Colloid Mill, MZ-130 (Fryma); capacity: 350 kg / hr) attached to a top loading hopper. The mill operation was 18 ° C. and 30 ° C. at a 1 R gap setting, chosen to achieve a particle size of less than 100 microns. All 45 kg seeds were ground within 10 minutes.

Homogenization and solids removal The resulting slurry is pumped into a knife in-line homogenizer (Dispax Reactor® DR 3-6 / A, IKA® Works, Inc.) at a rate of about 7 L / min. Sent by. After centrifugation to an operating speed of 3250 rpm, the output slurry was directly subjected to decantation centrifugation (NX-314B-31, Alfa-Laval). Within 25 minutes, approximately 160 kg of seed grind slurry was decanted. A Watson-Marlow (Model 704) peristaltic pump was used for this stage of slurry transfer.

Oil Body Separation Oil body fractions can be separated by three-phase separation and self-cleaning bowl and removable ring dam series; maximum capacity: 83L / min; ring dam: 69mm equipped with disk stack centrifugal separator (SA 7 , Westfalia). The operating speed was ~ 8520 rpm. Using a Watson-Marlow (Model 704) peristaltic pump, the decanted liquid phase (DL) was pumped to the centrifuge after being brought to operating speed. This decanted the liquid phase into a heavy phase (HP1) containing water and soluble seed protein, and a light phase (LP1) containing oil bodies. The oil body fraction obtained after one centrifugation is called the unwashed oil body preparation. The unwashed oil body is then passed through a static in-line mixer with mixing with 25 mM NaH ₂ PO ₄ (pH 7) buffer (35 ° C., 4 L / min) to a second disk stack centrifuge separator ( SA 7, Westfalia); maximum capacity: 83L / min; ring dam: 73mm. The operating speed was ~ 8520 rpm. The separated light phase (LP2) containing the oil body was then passed through another static in-line mixer while mixing with 50 mM NaH ₂ P0 ₄ buffer at pH 8 (35 ° C., 4 L / min). To a disk stack centrifuge separator (SA 7, Westfalia); maximum capacity: 83 L / min; ring dam: 75 mm. The operating speed was ~ 8520 rpm. The entire procedure was performed at room temperature. All the preparations obtained after the second separation are called washed oil body preparations. After three washes, most of the contaminating soluble seed protein was removed. If oil bodies are used in cosmetic formulations, 0.1% Neolone 950 and 0.1% glycacil L may be added as preservatives.

Example 2
Partitioning of clobetasol propionate into washed safflower oil bodies Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Clobetasol propionate (CP) (supplier-Sigma) was weighed (12 mg-30 mg) in a clean and dry 16 × 100 mm screw cap Pyrex tube and mixed with 300 μl isopropanol followed by 200 mg safflower oil. The combined samples were vortexed and subsequently incubated at 34 ° C. for 20 minutes. After incubation, the sample was revortexed and then dried under nitrogen for 20 minutes to remove isopropanol. Add 1 ml of washed high dry weight oil body to the CP / benivena oil mixture at room temperature, briefly centrifuge, pellet the contents, allow thorough mixing of the contents, vortex, Further incubation in an airtight tube at 34 ° C. to 42 ° C. overnight allowed the uptake of CP and safflower oil into the oil body. Hexane extraction was used to determine the amount of CP present in free oil (free oil is defined as the oil not contained in the oil body. Intact oil bodies are mostly hexane alone. All the hexane extraction of added oil bodies is resistant to extraction by hexane, but the damage caused to intact oil bodies by hexane is free of free oil obtained from unadded oil bodies. Note that corrections must be made through corrections that use values). Add 3 ml of hexane to the tube and shake the tube 32 times to mix. Samples are centrifuged in a swinging bucket clinical centrifuge at 3220 × g for 1 minute to separate hexane from the oil body-water phase. Note that free oil remains in the hexane layer (upper layer). The hexane layer is transferred to another tube and the hexane extraction is repeated for the remaining CP / oil body mixture. Once the majority of the solvent from the second extraction is added to the tube containing the first extract, the hexane-containing tube is transferred to a heating block. Hexane is at least 1.5 hours, while heating the block to 40 ° C. to 45 ° C., evaporated by subjecting the tubes to a gentle stream of high purity N ₂ gas. Once free oil was removed from the CP / oil body fraction, an analysis of the total remaining oil was performed. Add remaining whole oil in intact oil body by adding 4 ml of 3: 2 hexane: isopropanol solution (HIP) and mix until all oil is dissolved in HIP solvent (about 10-20 seconds) Measured by shaking vigorously. This was followed by the addition of 2.5 ml of 6.67% Na ₂ SO ₄ (w / v) to the tube and the tube was shaken for an additional 10 seconds. Phase separation is facilitated by centrifugation at 3220 × g for 2 minutes in a swinging bucket clinical centrifuge. The organic or upper phase is transferred to a second test tube using a Pasteur pipette, avoiding aqueous phase migration. 3 ml of 7: 2 HIP solution is added to the original tube containing the aqueous phase and the tube is shaken for 10 seconds. The tube is centrifuged for 2 minutes at 3220 × g and the upper phase is combined with the organic phase recovered in the first HIP extraction. The 7: 2 HIP extraction process was repeated. While heating in a dry block heater (40 ° C. to 45 ° C.), the solvent was evaporated from the liquid extract by subjecting the tube containing the combined organic phases to a gentle stream of compressed N ₂ gas. The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and only the extracted lipid remains. The amount of CP present in both the free oil fraction and the total oil fraction was determined by high performance liquid chromatography (HPLC) at a wavelength of 240 nm, and then an HPLC standard curve prepared with a known amount of CP and Compared. By comparing the amount of CP in both the free oil fraction and the total oil fraction, it was determined that an average of 94.7% of the added clobetasol propionate was incorporated into the intact oil body. The level of clobetasol propionate incorporated into the oil body was 0.316% as a percentage of dry weight. Cito-unguator lab mixer (Gako Konietzko) has been found to be particularly efficient in mixing oil bodies and oils when adding larger volumes of oil bodies and thus facilitating efficient additions .

Example 3
Distribution of retinoic acid to washed safflower oil bodies Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Retinoic acid (RA) (supplier-Sigma) was weighed (1 mg-8 mg) in a clean and dry 16 × 100 mm screw cap Pyrex tube and mixed with 3 ml of isopropanol. Add safflower oil so that there is no more than 5 mg RA per gram safflower oil. The combined samples were vortexed and placed in a 40 ° C. to 45 ° C. heating block and then dried under a steady stream of nitrogen until the isopropanol was removed (approximately 0.5 hour to 1 hour). Next, 4 ml to 5 ml of washed high dry weight oil body per gram of safflower oil used to solubilize RA is added to the RA / safflower oil mixture. Note that the RA / safflower oil mixture was kept in the heating block until just prior to the addition of the oil body. This oil body mixture is briefly centrifuged to pellet the contents, allowing thorough mixing of the contents, vortexing, and incubating overnight at 34 ° C to 37 ° C in an airtight tube. Enables the incorporation of RA and safflower oil into the oil body. Hexane extraction was used to determine the amount of unincorporated RA still dissolved in the free oil (free oil is defined as the oil not contained in the oil body. Oil bodies are mostly resistant to extraction with hexane alone, but all hexane extractions of added oil bodies are free of added oil for damage done to intact oil bodies by hexane. Note that corrections must be made through corrections using free oil values obtained from the body). Add 3 ml of hexane to the tube and shake the tube 32 times to mix. Samples are centrifuged in a swinging bucket clinical centrifuge at 3220 × g for 1 minute to separate hexane from the oil body-water phase. Note that free oil remains in the hexane layer (upper layer). The hexane layer is transferred to another tube and the hexane extraction is repeated for the remaining RA / oil body mixture. Once the majority of the solvent from the second extraction is added to the tube containing the first extract, the hexane-containing tube is transferred to a heating block. Hexane is at least 1.5 hours, while heating the block to 40 ° C. to 45 ° C., evaporated by subjecting the tubes to a gentle stream of high purity N ₂ gas. Once free oil was removed from the RA / oil body fraction, an analysis of the total remaining oil was performed. Add remaining whole oil in intact oil body by adding 4 ml of 3: 2 hexane: isopropanol solution (HIP) and mix until all oil is dissolved in HIP solvent (about 10-20 seconds) Measured by shaking vigorously. This was followed by the addition of 2.5 ml of 6.67% Na ₂ SO ₄ (w / v) to the tube and the tube was shaken for an additional 10 seconds. Phase separation is facilitated by centrifugation at 3220 × g for 2 minutes in a swinging bucket clinical centrifuge. The organic or upper phase is transferred to a second test tube using a Pasteur pipette, avoiding aqueous phase migration. 3 ml of 7: 2 HIP solution is added to the original tube containing the aqueous phase and the tube is shaken for 10 seconds. The tube is centrifuged for 2 minutes at 3220 × g and the upper phase is combined with the organic phase recovered in the first HIP extraction. The 7: 2 HIP extraction process was repeated. While heating in a dry block heater (40 ° C. to 45 ° C.), the solvent was evaporated from the liquid extract by subjecting the tube containing the combined organic phases to a gentle stream of compressed N ₂ gas. The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and that only the extracted lipid remains. The amount of RA present in both the free oil fraction and the total oil fraction is calculated by measuring absorption using a spectrophotometer at a wavelength of 380 nm and then prepared with a known amount of RA Compared to a standard curve. By comparing the amount of RA collected from the total oil fraction with that added to the oil body, it was determined that an average of 94.72% of the added RA was incorporated into the intact oil body. The level of RA incorporated into the oil body was 0.195% as a percentage of dry weight. Cito-unguator lab mixer (Gako Konietzko) has been found to be particularly efficient in mixing oil bodies and oils when adding larger volumes of oil bodies and thus facilitating efficient additions .

Example 4
Distributing Disranol to Washed Safflower Oil Body Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Disranol (supplier-Spectrum) was weighed (1 mg-30 mg) in a clean and dry 16 × 100 mm screw cap Pyrex tube and mixed with 500 μl of chloroform. Add safflower oil so that there is no more than 9 mg of disranol per gram of safflower oil. The combined samples were vortexed and placed in a 40 ° C. to 45 ° C. heating block and then dried under a steady stream of nitrogen until the chloroform was removed (remixing the mixture from time to time for about 1 hour to 2 hours). Next, 4 ml to 5 ml of washed high dry weight oil body containing 0.2% L-ascorbic acid (supplier-Sigma) per gram of safflower oil used to solubilize the disranol, was added to the dithranol / safflower oil. Add to mixture. This oil body mixture is briefly centrifuged to pellet the contents, allowing thorough mixing of the contents, vortexing, and incubating overnight at 34 ° C to 37 ° C in an airtight tube. This enables the incorporation of disulanol and safflower oil into the oil body. The oil body is washed once with an equal volume of 50 mM phosphoric acid containing 0.2% ascorbic acid, pH 8.0 (Note: unincorporated disranol pellets during the wash. Disranol is not soluble in hexane and Thus, hexane extraction does not remove unincorporated disulanol.), Remove oil body from upper phase and place in fresh tube. Hexane extraction was used to determine the amount of free oil, indicating the efficiency of the oil carrier addition (free oil is defined as the oil not contained in the oil body. All hexane extraction of the added oil body is obtained from the unadded oil body for damage done to the intact oil body by hexane, although the portion is resistant to extraction with hexane alone Note that corrections must be made through corrections using free oil values). Add 3 ml of hexane to the tube and shake the tube 32 times to mix. Samples are centrifuged in a swinging bucket clinical centrifuge at 3220 × g for 1 minute to separate hexane from the oil body-water phase. Note that free oil remains in the hexane layer (upper layer). The hexane layer is transferred to another tube and the hexane extraction is repeated for the remaining dithranol / oil body mixture. Once the majority of the solvent from the second extraction is added to the tube containing the first extract, the hexane-containing tube is transferred to a heating block. Hexane is at least 1.5 hours, while heating the block to 40 ° C. to 45 ° C., evaporated by subjecting the tubes to a gentle stream of high purity N ₂ gas. Once free oil was removed from the dithranol / oil body fraction, an analysis of the total remaining oil was performed. The remaining total oil in the intact oil body was measured by adding 3 ml of chloroform and shaking vigorously to mix until all the oil was dissolved in the solvent (about 10-20 seconds). Phase separation is facilitated by centrifugation at 3220 × g for 1 minute in a swinging bucket clinical centrifuge. The organic or upper phase is transferred to a second test tube using a Pasteur pipette, avoiding aqueous phase migration. 3 ml of chloroform is added to the original tube containing the aqueous phase and the tube is shaken for 10 seconds. The tube is centrifuged for 1 minute at 3220 × g and the upper phase is combined with the organic phase recovered in the first chloroform extraction. 3 ml of 7: 2 hexane: isopropanol solution (HIP) is added to the original tube containing the aqueous phase and the tube is shaken for 10 seconds. The tube is then centrifuged for 2 minutes at 3220 × g and the upper phase is combined with the lower chloroform phase obtained in the first two steps. While heating in a dry block heater (40 ° C. to 45 ° C.), the solvent was evaporated from the liquid extract by subjecting the tube containing the combined organic phases to a gentle stream of compressed N ₂ gas. The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and only the extracted lipid remains. The amount of disranol present in both the free oil fraction and the total oil fraction is calculated by measuring the absorption using a spectrophotometer at a wavelength of 376 nm and then prepared with a known amount of disranol Compared to a standard curve. By comparing the amount of dithranol recovered from the total oil fraction with that added to the oil body, it was determined that an average of 97.3% of the added disranol was taken up into the intact oil body. The level of disranol incorporated into the oil body was 0.253% as a percentage of dry weight. Cito-unguator lab mixer (Gako Konietzko) has been found to be particularly efficient in mixing oil bodies and oils when adding larger volumes of oil bodies and thus facilitating efficient additions .

Example 5
Distribution of Diclofenac into Washed Safflower Oil Body Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Diclofenac (supplier-Sigma) was weighed (1 mg-300 mg) in a clean and dry 16 × 100 mm screw cap Pyrex tube, with 10 ml ethanol and 3 volumes phosphate buffer (with 0.1% Neolone 950, 50 mM monobasic sodium phosphate, pH 8.0) was added to the ethanol / diclofenac mixture and mixed. 1 g of oil body is added to the buffered ethanol mixture at room temperature and the sample is incubated overnight at 34 ° C. to 37 ° C. in an airtight tube to allow uptake of diclofenac into the oil body. Oil bodies are centrifuged at 3220 × g for 10 minutes to separate them from buffer containing ethanol and possibly unincorporated diclofenac. Remove the buffer and wash the oil body twice with 50 mM phosphoric acid, pH 8.0 containing 0.1% Neolone 950. Chloroform :: methanol (2: 1) extraction was used to determine the amount of diclofenac contained in the oil extracted from the oil bodies. 3 ml of chloroform :: methanol is then added to the tube and the tube is shaken vigorously to mix. The sample is centrifuged in a swinging bucket clinical centrifuge at 3220 × g for 1 minute to separate the solvent from the oil body-water phase. The solvent layer is transferred to another tube and the extraction is repeated two more times for the remaining diclofenac / oil body mixture. The second extract and the third extract were added to the tube containing the first extract. The tube containing the extract is transferred to a heating block. The solvent is at least 1.5 hours, while heating the block to 40 ° C. to 45 ° C., evaporated by subjecting the tubes to a gentle stream of high purity N ₂ gas. The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and only the extracted lipid remains. The amount of diclofenac present in the total oil fraction was calculated by measuring the absorption using a spectrophotometer at a wavelength of 320 m and then compared with a standard curve prepared with a known amount of diclofenac. By comparing the amount of diclofenac recovered from the total oil fraction with that added to the oil body, it was determined that an average of 43.9% of the added diclofenac was incorporated into the intact oil body. As a percentage of dry weight, the level of diclofenac incorporated into the oil body was 1.36%.

Example 6
Distribution of Tetracaine into Washed Safflower Oil Body Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Tetracaine free base (supplier-Sigma) was weighed (1 mg-200 mg) in a clean and dry 16 × 100 mm screw cap Pyrex test tube and mixed with 1 ml isopropanol and then 1 g safflower oil. Next, 3 g to 5 g of washed high dry weight oil body is added to the tetracaine / oil mixture at room temperature, the sample is thoroughly mixed, and overnight at 34 ° C. to 37 ° C. in an airtight tube. Incubate to allow incorporation of tetracaine into the oil body. Using hexane extraction, the amount of free oil was determined to determine the amount of tetracaine still dissolved in the free oil (free oil as oil not contained in the oil body). Intact oil bodies are mostly resistant to extraction with hexane alone, but all hexane extraction of the added oil body is about damage done to intact oil bodies by hexane. Note that corrections must be made through corrections using free oil values obtained from unadded oil bodies). After 3 ml of hexane is added to the tube, the tube is shaken 32 times to mix. Samples are centrifuged in a swinging bucket clinical centrifuge at 3220 × g for 1 minute to separate hexane from the oil body-water phase. Note that free oil remains in the hexane layer (upper layer). The hexane layer is transferred to another tube and the hexane extraction is repeated for the remaining tetracaine / oil body mixture. Once the majority of the solvent from the second extraction is added to the tube containing the first extract, the hexane-containing tube is transferred to a heating block. Hexane is at least 1.5 hours, while heating the block to 40 ° C. to 45 ° C., evaporated by subjecting the tubes to a gentle stream of high purity N ₂ gas. Once free oil was removed from the tetracaine / oil body fraction, an analysis of the total remaining oil was performed. Add remaining whole oil in intact oil body by adding 4 ml of 3: 2 hexane: isopropanol solution (HIP) and mix until all oil is dissolved in HIP solvent (about 10-20 seconds) Measured by shaking vigorously. This was followed by the addition of 2.5 ml of 6.67% Na ₂ SO ₄ (w / v) to the tube and the tube was shaken for an additional 10 seconds. Phase separation is facilitated by centrifugation at 3220 × g for 2 minutes in a swinging bucket clinical centrifuge. The organic or upper phase is transferred to a second test tube using a Pasteur pipette, avoiding aqueous phase migration. 3 ml of 7: 2 HIP solution is added to the original tube containing the aqueous phase and the tube is shaken for 10 seconds. The tube is centrifuged for 2 minutes at 3220 × g and the upper phase is combined with the organic phase recovered in the first HIP extraction. The 7: 2 HIP extraction process was repeated. The solvent was evaporated by subjecting the tube to a gentle stream of compressed N ₂ gas while heating in a dry block heater for at least 1.5 hours (40 ° C. to 45 ° C.). The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and that only the extracted lipid remains. The amount of tetracaine present in the total oil fraction was calculated by measuring the absorption using a spectrophotometer at a wavelength of 338 nm and then compared to a standard curve prepared with a known amount of tetracaine. . By comparing the amount of dithranol recovered from the total oil fraction and that added to the oil body, it was determined that an average of 90.8% of the added tetracaine was incorporated into the intact oil body. The level of tetracaine incorporated into the oil body was 2.43% as a percentage of dry weight. Cito-unguator lab mixer (Gako Konietzko) has been found to be particularly efficient in mixing oil bodies and oils when adding larger volumes of oil bodies and thus facilitating efficient additions .

Example 7
Distribution of Phosphatidylcholine into Washed Safflower Oil Body Washed safflower oil bodies were prepared as described in Example 1. Oil bodies were stored with 0.1% Neolone 950 and 0.1% glycacil L. Phosphatidylcholine (PC, supplier-Sigma) was weighed (1 mg-300 mg) in a clean and dry 16 × 100 mm screw cap Pyrex tube. A fluorescent tracer (phosphatidic acid, supplier—Molecular Probes) was added to the PC. The amount used was 0.05% to 0.25% PC weight of 1 mg / ml tracer solution (resuspended according to manufacturer's instructions). 200 μl of isopropanol was added every 10 mg of PC to dissolve the PC / tracer mixture. Buffer _{(50mM NaH 2 PO 4, pH} 8.0), was added to using the same buffer as the oil bodies are made therefrom _{(50mM NaH 2 PO 4, pH} 8.0) isopropanol mixture. The volume was equivalent to the volume of the added oil body. The PC / tracer / isopropanol / buffer mixture was sonicated for 15-30 seconds. The solvent was evaporated using a steady stream of nitrogen with gentle heating (42 ° C.) of the sample for 15 minutes. The oil bodies were incubated at 37 ° C. for hours to days in airtight tubes. The oil body was washed twice with buffer to remove unincorporated PC. Both the wash and oil body fractions were evaluated for the amount of fluorescence after extraction with hexane using the entire oil extraction procedure. To add the entire oil extraction, add 4 ml of 3: 2 hexane: isopropanol solution (HIP) to the measured sample volume and mix until all the oil is dissolved in the HIP solvent (about 10-20 seconds) By vigorous shaking. 2.5 ml of 6.67% Na ₂ SO ₄ (w / v) was added to the tube and the tube was shaken for an additional 10 seconds. Phase separation is facilitated by centrifugation at 3220 × g for 2 minutes in a swinging bucket clinical centrifuge. The organic or upper phase is transferred to a second test tube using a Pasteur pipette, avoiding aqueous phase migration. 3 ml of 7: 2 HIP solution was added to the original tube containing the aqueous phase and the tube was shaken for 10 seconds. The tube is centrifuged for 2 minutes at 3220 × g and the upper phase is combined with the organic phase recovered in the first HIP extraction. The 7: 2 HIP extraction process was repeated. While heating in a dry block heater (40 ° C. to 45 ° C.), the solvent was evaporated from the lipid extract by subjecting the tube containing the combined organic phases to a gentle stream of compressed N ₂ gas. The tube is weighed after 1 hour and then every 15 minutes thereafter. If two consecutive weights are the same (± 0.0001 g), assume that all volatile components have evaporated and only the extracted lipid remains. A known amount of isopropanol was added to the extraction fraction and the sample was quantified using a fluorescence spectrophotometer. By comparing the amount of fluorescence recovered from the oil body with that added to the oil body as a tracer, an amount of PC equivalent to an average of 0.12% of the dry weight of the oil body can be added to the oil body film. It was determined. Stearylamine is derived from stearic acid (from beef tallow) and ammonia. This is a positive charge inducer that has been used to modify the charge of PC liposomes (Moncelli et al. (1994) Biophys. J. 66: 1969-1980). If stearylamine (Sigma) is included in the PC / tracer mixture at about 30% of the amount of PC used for the addition, the amount of PC can be added to the oil body membrane. It was equivalent to an average of 1.25% by weight.

  Although the present invention has been described with reference to what are presently considered to be the preferred embodiments, it is understood that the invention is not limited to the disclosed embodiments. On the contrary, the invention is intended to cover various modifications and equivalent mechanisms falling within the spirit and scope of the appended claims.

  All publications, patents, and patent applications are as if each individual publication, patent, or patent application was shown to be specifically and independently incorporated by reference in their entirety, The entirety of which is incorporated herein by reference.

Claims (34)

A method of distributing an active agent to an oil body comprising the following steps:
a) dissolving the active agent in the first solvent;
b) mixing the dissolved active agent with a second solvent to obtain a mixture of a first solvent and a second solvent comprising the active agent; and
c) contacting the active agent with the oil body by contacting the mixture of the first solvent and the second solvent with the oil body;   2. The method of claim 1, wherein the active agent is not partitioned into the oil body when contacted with the oil body in the absence of a solvent or when the active agent is dissolved in the first solvent alone.   3. The method of claim 2, wherein the active agent is insoluble in water.   The method of claim 2, wherein the active agent is insoluble in the second solvent.   The method of claim 1, wherein the amount of active agent distributed in the oil body ranges from about 0.0001% to 50% (w / v).   The method of claim 1, wherein the amount of active agent distributed in the oil body ranges from about 0.1% to 20% (w / v).   The method of claim 1, wherein the amount of active agent distributed in the oil body ranges from about 0.1% to 10% (w / v).   The process of claim 1 wherein the efficiency of partitioning the activity into the intact oil body is in the range of about 10% to 99%.   The method of claim 1 wherein the efficiency of partitioning the activity into intact oil bodies is in the range of about 50% to 99%.   The method of claim 1 wherein the efficiency of partitioning activity into intact oil bodies is in the range of about 90% to 99%.   2. The method of claim 1, wherein the active agent is selected from the group of active agents consisting of hydrophobic molecules and amphiphilic molecules.   12. The method of claim 11, wherein the hydrophobic molecule is selected from the group consisting of clobetasol propionate, diclofenac, disranol, retinoic acid, lidocaine, clindamycin, benzoyl peroxide, and cyclosporin A.   12. The method of claim 11, wherein the hydrophobic molecule has a log P value in the range of about 0-8.   12. The method of claim 11, wherein the hydrophobic molecule has a log P value in the range of about 2-7.   12. The method of claim 11, wherein the hydrophobic molecule has a log P value in the range of about 3-7.   12. The method of claim 11, wherein the amphiphilic molecule is selected from the group consisting of amphotericin B, phosphatidylcholine, tetracaine, and actinomycin D.   12. The method of claim 11, wherein the amphiphilic molecule has an HLB value in the range of about 1-14.   12. The method of claim 11, wherein the amphiphilic molecule has an HLB value in the range of about 4-10.   12. The method of claim 11, wherein the amphiphilic molecule has an HLB value in the range of about 6-8.   The method of claim 1, wherein the first solvent is not compatible with the oil body or is undesirable in the final product.   2. The method according to claim 1, wherein the first solvent is an organic solvent.   The first solvent is a solvent consisting of alcohol, aliphatic hydrocarbon, aromatic hydrocarbon, chlorinated hydrocarbon, glycol, glycol ether, and acetates, esters, ethers, ketones, oils, lipids, and fatty acids thereof; 2. The method of claim 1, wherein the method is selected from the group.   24. The method of claim 22, wherein the first solvent is an alcohol or chlorinated hydrocarbon.   24. The method of claim 22, wherein the first solvent is selected from the group consisting of isopropanol, ethanol, and chloroform.   2. The method of claim 1, wherein the second solvent is selected from the group of solvents consisting of water, aqueous buffer, oil, fatty acid, and lipid.   26. The method of claim 25, wherein the aqueous buffer is selected from the group consisting of 50 mM sodium phosphate monobasic, pH 8.0 and 25 mM sodium bicarbonate, pH 8.3.   26. The method of claim 25, wherein the oil is safflower oil.   The method of claim 1, wherein the first solvent is substantially removed after mixing with the second solvent.   27. The method of claim 26, wherein the first solvent is substantially removed by evaporation or the volume is substantially reduced by dilution.   28. The method of claim 27, wherein the method of evaporation is exposing the sample to a stream of nitrogen.   2. The method of claim 1, wherein the oil body is obtained from cells comprising an oil body or an oil body-like organelle.   30. The method of claim 29, wherein the cells include animal cells, plant cells, fungal cells, yeast cells, bacterial cells, and algal cells.   32. The method of claim 30, wherein the plant cells include pollen, spores, seeds, and cells from plant vegetative organs.   Plant seeds are rapeseed (Brassica spp.), Soybean (Glycine max), sunflower (Helianthus annuus), oil palm (Elaeis guineeis), cottonseed (Gossium) (Gossypium spp.)), Groundnut (Arachis hypogaea), coconut (Cocus nucifera), castor (Ricinus communis), olive (Olea spp.), Safflower (Carthas) Tintrius (Carthamus tinctorius), mustard (Brassica spp.) And Sinapis alba, coriander (Coriandrum sativum), pumpkin (Cucurbita maxima), flax / ama Linum usitatis simum), Brazil nuts (Bertholletia excelsa), jojoba (Simmondsia chinensis), corn (Zea mays), hamana (Crambe abyssinica), and Lucola el 32. A method according to claim 31 obtained from the group of plant species consisting of (Eruca sativa)).